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Fig. 10. MYC downregulates mammary expression of Cav1 in a developmental
stage-specific manner. (A) Northern hybridization analysis. Total RNA was
extracted from the mammary glands of MTB and MTB/TOM mice induced with
doxycycline from D6.5-D9.5 of pregnancy and sacrificed at D9.5 or D12.5 of
pregnancy, as well as from MTB and MTB/TOM mice induced from D12.5-D15.5 and
sacrificed at D15.5 or D18.5 of pregnancy. Total RNA was subject to gel
electrophoresis and blotted to nitrocellulose membranes. Blots were probed
with radiolabeled cDNA fragments specific to the human MYC transgene,
as well as mouse Cav1, Cish and CK18. Ethidium bromide
staining of 28S RNA is shown as a loading control. (B) Western blot analysis
of Cav1 expression. Mammary protein lysates from MTB and MTB/TOM mice induced
from D6.5-D8.5 or D12.5-D14.5 of pregnancy were separated by SDS-PAGE and
blotted to nitrocellulose. FVB wild-type mammary lysates from virgin, D7.5,
D12.5 and D18.5 of pregnancy were included as controls. Blots were probed with
antibodies specific to Cav1 and ß-tubulin.
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